Effect Of VEGFR3–ASODN And NRP2-ASODN On Proliferationandapoptosis Of Gastric Cercinomacells

ObjectiveTo investigate and compare the influence of each blocking and thesame time blocking the gene expression of VEGFR3and NRP2on theproliferation and apoptosis of SGC-7901cells.Methods(1) Divide SGC-7901cell lines into three groups, as VEGFR3blocking group（V group）, NRP2blocking group（N group）and VEGFR+NRP2blocking group（T group）, each big group and control groupcontains blank, liposomes transfected group, no righteous chain transfectedgroup (NSODN group) and different concentration "antisense" chaintransfected group (ASODN group).(2) With the content and LipofectamineTM2000will sulfatedmodified VEGFR3-ASODN, NRP2-ASODN and no righteousoligonucleotide transfected into human cancer SGC-7901within the celllines, in the cart before the horse after a microscope transfected cells.(3) With RT-PCR method to detect the cells VEGFR3-mRNA and NRP2-mRNA expression level.(4) Use MTT method and flow cytometry art detection cellproliferation and apoptosis.(5) Compared the data of my research with the data from othermembers in our research group at the same period.Results(1) After24h of the transfection, observe each group of cells，underthe fluorescence microscope, we find that control and blank liposometransfected group were no fluorescence expression, NSODN transfectedgroup and ASODN transfected group can be obviously positions of greenfluorescent in the cytoplasm of stomach cancer cells.(2) In the Group V,the level of VEGFR3gene transcription ofVEGFR3-ASODN transfected cells was significantly lower than the blankcontrol, liposomes transfected group and VEGFR3-NSODN transfectedgroup. The difference has statistics significance（P<0.05）. In the GroupN,the level of NRP2gene transcription ofNRP2-ASODN transfected cells was significantly lower than the blankcontrol, liposomes transfected group and NRP2-NSODN transfected group.The difference has statistics significance (P<0.05). There were nostatistically significant differences between blank control group, liposomestransfected group and NSODN transfected group, in the two biggroups(F=0.32，P=0.5760). (3) The results of MTT test of each group and time points is that underthe same conditions, the inhibitory effect of VEGFR3-ASODN to theproliferation SGC-7901was more obvious than NRP2-ASODN. Theinhibition effect was most obvious when transfected VEGFR3-ASODNand NRP2-ASODN at the same time.The difference all has statisticssignificance（P<0.05）.(4) Flow cytometry art tests showed that, the apoptosis rate of theSGC-7901cells when transfected VEGFR3-ASODN was higher than theSGC-7901cells transfected with NRP2-ASODN. The apoptosis rate of theSGC-7901cells was highest when transfected VEGFR3-ASODN andNRP2-ASODN at the same time, The difference has statistics significance（P<0.05）.(5) In the same condition, the proliferation inhibition rate andapoptosis rate of the NRP1-ASODN transfected group of stomach cancercell is higher than NRP2-ASODN transfected group. The proliferationinhibition rate of the VEGFR2-ASODN transfected group andVEGFR3-ASODN transfected group of stomach cancer cell was similar,the apoptosis rate of the former is higher than the latter. Compared theVEGFR3-ASODN and NRP2-ASODN union transfected group with theVEGFR2-ASODN and NRP1-ASODN union transfected group, theproliferation inhibition rate of stomach cancer cells was similar, the cellapoptosis rate of former is higher than the latter. Conclusion(1) Through the research we can see that, both of VEGFR3-ASODNand NRP2-ASODN have certain effect.to cell proliferation and apoptosis ofthe cancer of stomach.(2) If compared the VEGFR3with NRP2, the inhibition effect ofproliferation and the promotion effect of apoptosis of SGC-7901whentransfected witn VEGFR3-ASODN was stronger than the SGC-7901cellwhen tranfected with NRP2-ASODN, and VEGFR3-ASODN andNRP2-ASODN also transfected stomach cancer cells, the inhibition effectof proliferation and the promotion effect of apoptosis of SGC-7901wasstrongest when tranfected with NRP2-ASODN and VEGFR3-ASODN atthe same time. So we assume that VEGFR3and NRP2play a synergy incell proliferation and apoptosis of gastric cancer.(3) NRP1-ASODN had harder cytostatic action and Promoteapoptosis role for gastric cancer cell than NRP2-ASODN. The strength ofthe inhibition proliferation to cancer of the stomach cells for theVEGFR2-ASODN was similar as the VEGFR3-ASODN, but the formerpromote better than the latter in the role of apoptosis. The strength of theinhibition proliferation to cancer of the stomach cells for the unitedtranfected VEGFR3-ASODN and NRP2-ASODN was similar as the unitedtranfected VEGFR2-ASODN and NRP2-ASODN. But the former played aweak role in promote apoptosis to the latter.